We seek to create an affordable and easily implemented proteomic tool in the form of a low cost "enzyme activity and immunodetection chip" for evaluation of enzyme activities and antigenic determinations at the level of 400 detections per cm2. Specific Aim 1 will focus on the validation and calibration of enzyme activity chemistries and antigen determination chemistries that are suited for reduction to a chip-based format at the 5 nanoliter scale per reaction. Specific Aim 2 will focus on the development of low cost microfluidics technology for activation of the enzyme/Immuno chips using minimal volumes of biological sample. We seek to conduct 400 detections using 10 to 100 microliters of biological fluid. Specific Aim 3 will focus on the scale down of the chemistries of Aim 1 to a chip based format where each reaction volume is 5 nanoliters and these reactions are activated with test fluids using the microfluidics technology developed in Aim 2. Preliminary studies have demonstrated the feasibility of activating thousands of individual 5 nanoliter reaction centers with a microliter of fluid sample. The Phase I activities will lead toward establishing chip chemistries and microfluidic technologies for a commercialized chip to aid in prostate cancer diagnostics.